Diversity, distribution, and sustainability of traditional medicinal plants in Kaski district, western Nepal.

Medicinal plants are the primary source of traditional healthcare systems in many rural areas mostly in developing countries. This study aimed to document and analyze the diversity, distribution, and sustainability of the traditional medicinal plants used by the Gurung people of the Sikles region in western Nepal. Ethnobotanical data were collected through focus group discussions and individual interviews, and analyzed using descriptive and inferential statistics. Prior informed consent was obtained before each interview. Quantitative ethnobotanical indices such as informant consensus factor, relative frequency of citation, and use values were also calculated. A possible association among these indices was tested using correlation analysis. A total of 115 wild medicinal plant species belonging to 106 genera and 71 families were documented. Asteraceae and Rosaceae were the dominant families whereas herbs were the most dominant life form. Roots were the most used plant part, paste was the most common method of preparation, and most of the medical formulations were taken orally. The highest number of medicinal plants were used to treat stomach disorders. The average informant consensus value of 0.79 indicates a high consensus among respondents in selecting medicinal plants. Lindera neesiana, Neopicrorhiza scrophulariiflora, Paris polyphylla, and Bergenia ciliata were found to be high-ranking medicinal plants based on the relative frequency of citation and use value. The genders did not affect medicinal plants' knowledge but age had a significant correlation. Most of the informants agreed that medicinal plants are under pressure due to overharvesting and a lack of proper forest management practices. The number of medicinal plants reported from the study area indicates that the Gurung people possess rich traditional knowledge, and the vegetation of the Sikles region constitutes rich diversity of medicinal plants.

Foods from the wild: Local knowledge, use pattern and distribution in Western Nepal.

Locally harvested wild edible plants (WEPs) provide food as well as cash income for indigenous peoples and local communities, and they are of great importance in ensuring local food security. However, their uses and availability are poorly documented. This study aimed to enumerate WEP diversity and status of WEPs in a part of the Annapurna Conservation Area, Sikles region, where the population is dominated by the Gurung community. Ethnobotanical data were collected using guided field walks, semi-structured interviews, and field observation. The informant consensus method was employed and group discussions were conducted for triangulation of the information. Free listing and identification tests were performed to assess the knowledge of the informants. Both descriptive statistics and quantitative ethnobotanical methods were used for data analysis. A total of 72 wild food species belonging to 46 families and 61 genera were reported from the study area. Asparagaceae and Rosaceae were the dominant families, and herbs were the dominant life form. Fruits (34 species) were the most frequently used plant parts, followed by young shoots (16 species). Most edible plants were consumed in summer and during rainy seasons. While the age and type of informants had an influence on the number of enumerated plants, gender did not. Key informants and people aged 30-45 reported more species than other groups of respondents. Most of the knowledge about the use of WEPs was acquired from parents and relatives. The consumption of these plants was attributed to diversifying cuisine, spicing staple food, nutri-medicinal values, and cultural practices. People perceived the availability of WEPs to be gradually decreasing. However, WEPs are still abundant and diverse in the study area, and knowledge on their use is well-preserved. These resources provide food and nutrients to local people and can also be a source of cash income. Therefore, the documented information on WEPs may serve as baseline data for further studies on nutritional values and provide guidelines for safe collection. The results also revealed that many wild species are under growing pressure from various anthropogenic factors, suggesting effective community engagement is required for their conservation.

Phytochemical Investigations and In Vitro Bioactivity Screening on Melia azedarach L. Leaves Extract from Nepal.

Melia azedarach is a common tree used in the traditional medicine of Nepal. In this work, leaves were considered as source of bioactive constituents and composition of methanol extract was evaluated and compared with starting plant material. Flavonoid glycosides and limonoids were identified and quantified by HPLC-DAD-MSn approaches in dried leaves and methanolic extract, while HPLC-APCI-MSn and GC/MS analysis were used to study phytosterol and lipid compositions. ß-Sitosterol and rutin were the most abundant constituents. HPLC-APCI-MSn and HPLC-DAD-MSn analysis revealed high levels of phytosterols and flavonoids in methanolic extract accounting 9.6 and 7.5 % on the dried weight, respectively. On the other hand, HPLC/MSn data revealed that limonoid constituents were in minor amount in the extract <0.1 %, compared with leaves (0.7 %) indicating that degradation occurred during extraction or concentration procedures. The methanol extract was subjected to different bioassays, and antioxidant activity was evaluated. Limited inhibitory activity on acetyl and butyryl cholinesterase, as well as on amylase were detected. Moreover, tyrosinase inhibition was significant resulting in 131.57±0.51 mg kojic acid equivalents/g of dried methanol extract, suggesting possible use of this M. azedarach extract in skin hyperpigmentation conditions. Moderate cytotoxic activity, with IC50 of 26.4 µg/mL was observed against human ovarian cancer cell lines (2008 cells). Our findings indicate that the Nepalese M. azedarach leaves can be considered as valuable starting material for the extraction of phenolics and phytosterols, yielding extracts with possible cosmetic and pharmaceutical applications.

An Integrated LC-ESI-MSn and High Resolution LC-ESI-QTOF Approach for the Identification of Phloroglucinols from Nepalese Hypericum japonicum.

Phloroglucinols are characteristic constituents of Hypericumjaponicum that are claimed to exert several bioactivities, such as anti-inflammatory, anti-depressant and anti-viral ones. Phloroglucinols are unstable compounds and their synthesis is challenging; thus, isolation from natural sources is still one of the main strategies for obtaining these constituents in purified form. Assessing the presence of phloroglucinols in plant materials can be of interest for compound isolation, and LC-MS approaches afford sensitivity and specificity in this regard. In this work, we combined data from quadrupole-time of flight (QTOF) and ion trap (IT) mass spectrometers in order to assess the presence of the phloroglucinols characteristic of H. japonicum and to elucidate their MS fragmentation pathways. The identified compounds present similar structures bearing the 1,3,5-trihydroxybenzene core with different substitutions, which, in constituents at higher MW, is linked to 3',3'-dimethyl-6'-oxo-phlorisobutyrophenone by a methylene bridge. Differences in MS2 spectra of the considered phloroglucinols are useful for compound identification and differentiation, and to perform dereplication studies. Overall, the proposed approach could be useful for the analysis of phloroglucinols in H. japonicum and other plant species.

In Vitro Antibacterial Activities of Methanolic Extracts of Fruits, Seeds, and Bark of Zanthoxylum armatum DC.

Crude methanol extracts of fruits, seeds, and bark of Zanthoxylum armatum were investigated in vitro for antimicrobial activities against 9 different bacterial strains: Bacillus subtilis, Enterococcus faecalis, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysenteriae, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Staphylococcus epidermidis using agar well diffusion method, and the MBC values were determined. Only 5 bacteria, i.e., Bacillus subtilis, Enterococcus faecalis, MRSA, Staphylococcus aureus, and Staphylococcus epidermidis exhibited antibacterial properties against the different extracts. The fruit and seed extracts showed activities against 5 bacteria, while the bark extract was active against 2 bacteria only (MRSA and Staphylococcus aureus). Staphylococcus aureus was found to be more susceptible for all the extracts compared to other strains. The maximum ZOI of 20.72 mm was produced by fruits (wild) and 18.10 mm (cultivated) against Staphylococcus aureus. Likewise, the least ZOI of 11.73 mm was produced by seeds (wild) and 11.29 mm (cultivated) against Escherichia faecalis. Similarly, the lowest MBC value of 0.78 mg/mL was obtained for fruit extracts against MRSA, 1.56 mg/mL for fruits, seeds, and bark extracts against Bacillus subtilis, MRSA, and Staphylococcus aureus, and highest value of 50 mg/mL for fruits and seeds extracts against S. epidermidis. The fruits, seeds, and bark extracts of Z. armatum exhibited remarkable antibacterial properties against different pathogenic bacteria causing several diseases, which suggests the potential use of this plant for treating different bacterial diseases such as skin infection, urinary tract infection, dental problems, diarrhea, and dysentery.

Himalayan Nettle Girardinia diversifolia as a Candidate Ingredient for Pharmaceutical and Nutraceutical Applications-Phytochemical Analysis and In Vitro Bioassays.

Girardinia diversifolia, also known as Himalayan nettle, is a perennial herb used in Nepal to make fiber as well as in traditional medicine for the treatment of several diseases. To date, phytochemical studies and biological assays on this plant are scarce. Thus, in the present work, the G. diversifolia extracts have been evaluated for their potential pharmaceutical, cosmetic and nutraceutical uses. For this purpose, detailed phytochemical analyses were performed, evidencing the presence of phytosterols, fatty acids, carotenoids, polyphenols and saponins. The most abundant secondary metabolites were ß- and γ-sitosterol (11 and 9% dw, respectively), and trans syringin (0.5 mg/g) was the most abundant phenolic. Fatty acids with an abundant portion of unsaturated derivatives (linoleic and linolenic acid at 22.0 and 9.7 mg/g respectively), vitamin C (2.9 mg/g) and vitamin B2 (0.12 mg/g) were also present. The antioxidant activity was moderate while a significant ability to inhibit acetylcholinesterase (AChE), butyrilcholinesterase (BuChE), tyrosinase, α-amylase and α-glucosidase was observed. A cytotoxic effect was observed on human ovarian, pancreatic and hepatic cancer cell lines. The effect in hepatocarcinoma cells was associated to a downregulation of the low-density lipoprotein receptor (LDLR), a pivotal regulator of cellular cholesterol homeostasis. These data show the potential usefulness of this species for possible applications in pharmaceuticals, nutraceuticals and cosmetics.

Total Phenolic, Flavonoid Contents, and Antioxidant Activities of Fruit, Seed, and Bark Extracts of Zanthoxylum armatum DC.

Natural antioxidants present in several medicinal plants are responsible for inhibiting the harmful effects of oxidative stress. These plants contain polyphenols and flavonoids that act as free radical scavengers and reduce oxidative stress and may be an alternative remedy to cure various harmful human diseases. This study aims to quantify the total phenolic and flavonoid contents (TPC and TFC) and antioxidant properties of methanol extracts of fruits, seeds, and bark of an important medicinal and aromatic plant, Zanthoxylum armatum collected from wild and cultivated populations in Nepal. TPC was determined by Folin-Ciocalteu colorimetric method using gallic acid as standard, and various concentrations of the extract solutions were measured at 760 nm. TFC was calculated by aluminum chloride colorimetric assay. Quercetin was used as standard, and the absorbance was measured at 510 nm. The antioxidant potential of the different extracts was estimated by DPPH free radical scavenging assay, and the absorbance was measured at 517 nm. The highest TPC value was 226.3 ± 1.14 mg GAE/g in wild fruits, and the lowest was 137.72 ± 4.21 mg GAE/g in cultivated seeds. Similarly, the highest TFC value was 135.17 ± 2.02 mg QE/g in cultivated fruits, and the lowest was 76.58 ± 4.18 mg QE/g in cultivated seeds. The extracts showed variable antioxidant properties. The fruits exhibited excellent antioxidant properties with IC50 values of 40.62 µg/mL and 45.62 µg/mL for cultivated and wild fruits, respectively. Similarly, the IC50 values of the bark were 63.39 µg/mL and 67.82 µg/mL, respectively, for cultivated and wild samples. And the least antioxidant capacity was shown by the seeds extract with IC50 values of 86.75 µg/mL and 94.49 µg/mL for wild and cultivated seeds, respectively. The IC50 value of the standard ascorbic acid was 36.22 µg/mL. Different extracts of Z. armatum contain considerable amount of phenols and flavonoids, including antioxidant properties, suggesting the potential use of this species in pharmacy and phytotherapy as a source of natural antioxidants.

Phytochemical Fingerprinting and In Vitro Bioassays of the Ethnomedicinal Fern Tectaria coadunata (J. Smith) C. Christensen from Central Nepal.

Tectaria coadunata, an ethnomedicinal fern used in Nepal to treat a large number of diseases, has been poorly studied with regard to its phytochemical composition and possible bioactivity. This study was performed with the aim of supporting traditional medicine as a new source of bioactive constituents. Phytochemical compositions of methanol extracts were determined by nuclear magnetic resonance (NMR), liquid chromatography-diode array detector-mass spectrophotometry (LC-DAD-MS), and liquid chromatography-fluorescence-mass spectrometry. Quali-quantitative data revealed large amount of procyanidins, mainly of the A-type, as well as eriodictyol-7-O-glucuronide and luteolin-7-O-glucoronide as main constituents. The antioxidant, cytotoxic, and inhibitory activity of five enzymes that are implicated in human diseases was evaluated for the extract and fractions. High free-radical scavenging activity in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays and inhibitory activities against cholinesterases and tyrosinase were observed. Furthermore, a moderate cytotoxic effect was observed on the 2008 and BxPC3 cell lines. Overall results showed potential usefulness of this fern as a source of phytochemicals for pharmaceutical uses.

LC-MSn and HR-MS characterization of secondary metabolites from Hypericum japonicum Thunb. ex Murray from Nepalese Himalayan region and assessment of cytotoxic effect and inhibition of NF-κB and AP-1 transcription factors in vitro.

Hypericum japonicum Thunb. ex Murray is traditionally used in Nepal to treat several diseases, among whom inflammation and acute pain. Although several secondary metabolites from the same Hypericum species have been already characterized and considered for their pharmacological use, an exhaustive phytochemical characterization of H. japonicum from Nepal is lacking, as well as the assessment of its potential pharmacological properties. Hence, the aims of this study were the characterization of a methanolic extract of H. japonicum (HJME) collected from the Northern region of Nepal by LC-MSn and UPLC-QTOF. The assessment of in vitro inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein 1 (AP-1) transcription factors and HJME's cytotoxic effect on human cell lines was performed to evaluate the potential use of this herb as a source of anti-inflammatory and cytotoxic lead compounds. Fifty-seven phytoconstituents were identified, being mainly flavonoids, phloroglucinols, phenolic acids and xanthones. Although compounds characteristic of H. japonicum were detected (quercetin, quercetin-7-O-α-l-rhamnoside, quercitrin and hyperoside), several others are here reported for the first time in this species. The results from bioassays indicated that HJME could significantly reduce the viability of human THP-1 cells (IC50 = 5.4 ±â€¯1.1 µg mL-1), showing the promising potential of HJME as anti-tumor agent. Furthermore, HJME significantly decreased the activation of both NF-κB and AP-1 at the concentration of 2 µg mL-1. Overall, these data suggest that H. japonicum from Nepal could be used as a source of potential natural anti-inflammatory and anti-tumor lead compounds.

Zanthoxylum armatum DC.: Current knowledge, gaps and opportunities in Nepal.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum armatum DC. possesses several medicinal properties and has been commonly used in different indigenous medicinal practices to cure several diseases because of its stomachic, carminative and anthelmintic properties. AIM: This review paper aims to provide an update on and analysis of information about the ecology, uses, phytochemistry, pharmacology, trade opportunities, policy gaps for the commercialization of this species forming a basis for further scientific innovations MATERIALS AND METHODS: Information was gathered through a search of different books, journals, articles, annual reports, proceedings and web-based materials. RESULT: Alkaloids, sterols, phenolics, lignins coumarins, terpenoids and flavonoids have been identified from leaves, fruits, stem, bark and seeds. Its trade value is also very high with its manifold applications in Ayurveda, allopathy, general pharmacy, and other industries. Antimicrobial, antiviral, antioxidant, anti-inflammatory, cytotoxic, hepato-protective, insecticidal/larvicidal effects are of particular relevance. CONCLUSION: It is one of the prioritized medicinal plants for economic development in Nepal. Owing to its diverse applications, the species can be developed as an important commodity for alleviation of poverty in rural areas. The various ethno-pharmacological applications of Zanthoxylum armatum have been verified by several related researches. More extensive study on the individual specific phyto-component can lead to novel innovations for the well-being of mankind.

Diversity of use and local knowledge of wild edible plant resources in Nepal.

BACKGROUND: Wild edible plants (WEP) provide staple and supplement foods, as well as cash income to local communities, thus favouring food security. However, WEP are largely ignored in land use planning and implementation, economic development, and biodiversity conservation. Moreover, WEP-related traditional knowledge is rapidly eroding. Therefore, we designed this study to fulfill a part of the knowledge gap by providing data on diversity, traditional knowledge, economic potential, and conservation value of WEP from Nepal. METHODS: The information was collected through focus group discussions and key informant interviews. Percentage of general utility of the plants among the study communities was evaluated using the Chi-square (χ(2)) test of homogeneity. High priority species were identified after consultation with the local stakeholders followed by scoring based on defined criteria. Pairwise ranking was used to assess ethnoecological knowledge to identify the threats to WEP. RESULTS: We documented 81 species belonging to Angiosperms (74), Pteridophytes (5), and Fungi (2). Most of the species were used as fruits (44 species) followed by vegetables (36). Almost half of the species (47%) were also used for purposes other than food. From the species with market value (37% of the total), 10 were identified as high priority species. Pairwise ranking revealed that WEP are threatened mostly by habitat destruction, land-use change and over-harvesting. Some of these plants are crop wild relatives and could thus be used for crop improvement. Interestingly, our study also revealed that young people who spend most of the time in the forest as herdsmen are particularly knowledgeable of wild fruit plants. CONCLUSION: We provide empirical evidence from a relatively large area of Nepal about diversity and status of WEP, as well as methodological insights about the proper knowledge holders to consult. Regarding the unique and important knowledge they have on WEP, young people should be included when recruiting participants to ethnobotanical studies or to any type of consultation about WEP. The habit of using wild edible plants is still alive and is a traditional culinary practice that demonstrates rich traditional knowledge of local people. WEP were found to be important for livelihood as well as showing great potential for crop improvement. Priority species should be promoted for income generation activities through sustainable collection and trade. Communities should engage in minimizing the threats to these valuable resources.